4.5 Article

Disruption of Leishmania flagellum attachment zone architecture causes flagellum loss

Journal

MOLECULAR MICROBIOLOGY
Volume -, Issue -, Pages -

Publisher

WILEY
DOI: 10.1111/mmi.15199

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The flagellar pocket, a cell membrane invagination at the base of the flagellum, is tightly associated with cytoskeletal structures in Leishmania and other kinetoplastid parasites. In this study, the localization and function of Leishmania orthologues of Trypanosoma brucei FAZ proteins were screened and analyzed. Deletion of two FAZ proteins, FAZ27 and FAZ34, in the flagellum resulted in reduced cell body size and flagellum loss in some cells. Moreover, distinct changes in cell shape were observed after generating the null mutant, demonstrating the parasite's ability to adapt to morphological perturbations resulting from gene deletion.
Leishmania are flagellated eukaryotic parasites that cause leishmaniasis and are closely related to the other kinetoplastid parasites such as Trypanosoma brucei. In all these parasites there is a cell membrane invagination at the base of the flagellum called the flagellar pocket, which is tightly associated with and sculpted by cytoskeletal structures including the flagellum attachment zone (FAZ). The FAZ is a complex interconnected structure linking the flagellum to the cell body and has critical roles in cell morphogenesis, function and pathogenicity. However, this structure varies dramatically in size and organisation between these different parasites, suggesting changes in protein localisation and function. Here, we screened the localisation and function of the Leishmania orthologues of T. brucei FAZ proteins identified in the genome-wide protein tagging project TrypTag. We identified 27 FAZ proteins and our deletion analysis showed that deletion of two FAZ proteins in the flagellum, FAZ27 and FAZ34 resulted in a reduction in cell body size, and flagellum loss in some cells. Furthermore, after null mutant generation, we observed distinct and reproducible changes to cell shape, demonstrating the ability of the parasite to adapt to morphological perturbations resulting from gene deletion. This process of adaptation has important implications for the study of Leishmania mutants.

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